DESCRIPTION (provided by investigator): Cholera, caused by Vibrio cholerae serogroups O1 and O139, is the most severe of diarrheal diseases and continues to cause significant morbidity and mortality throughout the developing world. Presently, it is reported in approximately 52 countries worldwide. During our first study, we carried out biweekly clinical and environmental surveillance in four rural areas of Bangladesh. Analysis of our accumulated clinical and environmental data identified factors that are significantly predictive for occurrence of cholera outbreaks, including water temperature, pH, and conductivity of surface water, along with concentrations of plankton and cholera toxin- producing bacteria in surface waters. A significant finding was that zooplankton, namely copepods, serve as an effective host for V. cholerae. By tracking their phytoplankton food source, using satellite monitoring of chlorophyll a concentration, it is possible to estimate zooplankton populations. With our successes to date, we will conduct epidemiological and ecological surveillance in Chattak and Mathbaria, two widely separated geographical sites, where we have documented predictable seasonal cholera outbreaks: one in the spring and the other in the fall. Using improved assays, we will enumerate multiple morphologies and life stages of V. cholerae in environmental samples, including the viable but nonculturable form (VBNC). Characterization of genetic associations between clinical and environmental isolates and GIS surveillance will be conducted to establish key epidemiological relationships that have been difficult to document at the genomic level. To elucidate local variability in cholera outbreaks, we will investigate factors recognized from our previous study, as well as those identified by other investigators, but not yet included in our predictive model, namely microbial community dynamics, increased competence of V. cholerae attached to chitinous surfaces, and plankton community dynamics. Using environmental variables identified from our previous studies and incorporating the new variables that prove to be correlated with cholera cases, we will refine our model of cholera transmission with the intent to render it useful for predicting cholera outbreaks, thereby allowing early mobilization of preventative and treatment measures. The very recent availability of inexpensive oral cholera vaccines makes this objective even more relevant.